Studies On The Methods Of Priming Of Dendritic Cells With Hpv16e6e7

Cervical cancer is the second leading cause of cancer death among womenworldwide. Statistics indicated that there were about500000new cases of cervicalcancer worldwide every year, of which200000died. Traditional treatment of cervicalcancer such as surgery, radiotherapy and chemotherapy have their own limitations,and can not cure cervical cancer. Dendritic cell (DC) is the most powerful antigen-presenting cell in human body. Great progress has been achieved in the research onthe mechanism and clinical application of dendritic cell, and numerous studiesindicated that dendritic cell play an important role in tumor-specific immunity．Therefore, the priming of DC is a critical process in DC-mediated tumor-specificimmunotherapy. In our studies, different methods for DC priming by HPV16E6E7were explored, such as priming of DC with recombinant adenoviruses expressingHPV16E6E7or with the transfection of mRNA of HPV16E6E7transcribed in vitroby electroporation.Human papillomavirus (HPV) is a major etiological factor of cervical cancer, andabout99.7%of cervical cancers contain HPV, of which50%are associated withHPV16or HPV18. E6and E7proteins play major roles in the development of cervicalcancer and these two oncoproteins are expressed only in tumor tissues, not in normaltissues. We established in our experiments an in vitro system for the transcription ofHPV16E6E7mRNA and constructed recombinant adenoviruses expressingcodon-optimized HPV16E6E7gene which contained the transformation-deficiencymutations. After comparison of their priming efficiencies based on EGFP expression,our results indicated that priming of DC using recombinant adenoviruses was moreefficient than that using mRNA transfection in vitro.